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WO2017096114A1 - Procédés de détermination d'analytes dans des fluides - Google Patents

Procédés de détermination d'analytes dans des fluides Download PDF

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Publication number
WO2017096114A1
WO2017096114A1 PCT/US2016/064539 US2016064539W WO2017096114A1 WO 2017096114 A1 WO2017096114 A1 WO 2017096114A1 US 2016064539 W US2016064539 W US 2016064539W WO 2017096114 A1 WO2017096114 A1 WO 2017096114A1
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WO
WIPO (PCT)
Prior art keywords
metal
analyte
capture
less
containing particles
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/US2016/064539
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English (en)
Inventor
Marta Fernández SUÁREZ
Lisa Marshall
Martina MEDKOVA
Aaron Oppenheimer
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Daktari Diagnostics Inc
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Daktari Diagnostics Inc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Daktari Diagnostics Inc filed Critical Daktari Diagnostics Inc
Publication of WO2017096114A1 publication Critical patent/WO2017096114A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/28—Electrolytic cell components
    • G01N27/30—Electrodes, e.g. test electrodes; Half-cells
    • G01N27/327—Biochemical electrodes, e.g. electrical or mechanical details for in vitro measurements
    • G01N27/3275—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction
    • G01N27/3277—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction being a redox reaction, e.g. detection by cyclic voltammetry
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/28—Electrolytic cell components
    • G01N27/30—Electrodes, e.g. test electrodes; Half-cells
    • G01N27/327—Biochemical electrodes, e.g. electrical or mechanical details for in vitro measurements
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/06—Investigating concentration of particle suspensions
    • G01N15/0656—Investigating concentration of particle suspensions using electric, e.g. electrostatic methods or magnetic methods
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/28—Electrolytic cell components
    • G01N27/30—Electrodes, e.g. test electrodes; Half-cells
    • G01N27/327—Biochemical electrodes, e.g. electrical or mechanical details for in vitro measurements
    • G01N27/3271—Amperometric enzyme electrodes for analytes in body fluids, e.g. glucose in blood
    • G01N27/3273—Devices therefor, e.g. test element readers, circuitry
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/28—Electrolytic cell components
    • G01N27/30—Electrodes, e.g. test electrodes; Half-cells
    • G01N27/327—Biochemical electrodes, e.g. electrical or mechanical details for in vitro measurements
    • G01N27/3275—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction
    • G01N27/3278—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction involving nanosized elements, e.g. nanogaps or nanoparticles
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/416—Systems
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54313—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
    • G01N33/54326—Magnetic particles
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54366—Apparatus specially adapted for solid-phase testing
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/0098—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor involving analyte bound to insoluble magnetic carrier, e.g. using magnetic separation
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/01—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials specially adapted for biological cells, e.g. blood cells
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2458/00—Labels used in chemical analysis of biological material
    • G01N2458/30—Electrochemically active labels

Definitions

  • the concentration of the electrolyte is less than 1.0 M, less than 0.8 M, less than 0.6 M, less than 0.4 M, less than 0.2 M, or less than 0.1 M after adding the electrolyte to the fluid.
  • the electrolyte does not remove the silver particle from the bound complex upon introduction of the electrolyte.
  • at least 90%, at least 95%, or at least 99% of the silver particles in the bound complex are not removed from the bound complex upon introduction of the electrolyte.
  • FIG. 1C is a schematic drawing of a method for isolating an analyte-containing biological particle, according to one set of embodiments
  • a plurality of capture structures may be added to the fluid such that the analyte binds to at least a portion of the plurality of capture structures.
  • the analyte may attach or bind to a capture structure in any suitable manner.
  • a single analyte e.g., a single type of analyte, or a single number of analytes
  • more than one analyte e.g., more than one type of analyte, or more than one number of analytes
  • the plurality of capture substrates may include a portion of a surface of the fluidic device, such as a surface of a channel. In some embodiments, the plurality of capture substrates are areas on a surface of a channel that have been functionalized with a capture entity. In certain embodiments, the plurality of capture structures comprise a plurality of microfluidic posts.
  • a microfluidic device comprises a plurality of microfluidic posts (e.g., conjugated microfluidic posts), such that an analyte introduced into the microfluidic device binds to the plurality of microfluidic posts.
  • a plurality of metal- containing particles may then be added to the microfluidic device such that the metal- containing particle, analyte, and microfluidic post forms a bound complex.
  • suitable microfluidic devices and structures e.g., posts
  • a plurality of metal-containing particles may be added to the fluid such that the analyte attaches or binds to at least a portion of the plurality of metal- containing particles.
  • the analyte may attach or bind to a metal-containing particle in any suitable manner.
  • a single analyte e.g., a single type of analyte, or a single number of analyte
  • more than one analyte e.g., more than one type of analyte, or more than one number of analytes
  • the plurality of metal-containing particles may have any suitable average particle size. Although other sizes are possible, in some cases the average particle size of the metal- containing particles is relatively large (e.g., at least 100 nm, at least 200 nm) to increase the signal-to-noise ratio when using certain detection methods, as described in more detail below.
  • Combinations of the above-referenced ranges are also possible (e.g., at least 0.06 and less than or equal to 15 times, at least 0.06 and less than or equal to 6 times, at least 0.07 and less than or equal to 2 times, at least 0.2 and less than or equal to 2.5 times, at least 1 and less than or equal to 4 times, at least 2 and less than or equal to 6 times the average particle size of the plurality of capture structures).
  • Other ranges are also possible.
  • the average particle sizes used may be those prior to forming a complex with the analyte (e.g., a capture structure-analyte-metal- containing particle complex). In some embodiments, the average particle sizes may be those prior to exposure of the particles to the electrolyte used in a detection step.
  • relatively large metal-containing particles may be used.
  • relatively larger metal-containing particles may result in an increased amplification of the signal during detection, which may increase the sensitivity of the methods described herein as compared to traditional quantification methods, which utilize relatively smaller (e.g., less than 40 nm) metal-containing particles.
  • the method comprises exposing the bound complex to an electrolyte (e.g., an electrolyte that may facilitate detection of the analyte).
  • the exposing step does not release the analyte from the metal-containing particle and/or the capture structure.
  • bound complex 150 may undergo exposing step 165.
  • the exposing step comprises exposing bound complex 150 to electrolyte 160, e.g., by adding the electrolyte to a fluid containing the bound complexes.
  • exposing step 165 may occur simultaneously with adding step 115 and/or adding step 135.
  • Simple screening tests can be employed to help select an electrolyte.
  • One simple screen test includes incubating the bound complex with the electrolyte for five minutes and then removing the electrolyte.
  • the amount of metal-containing particles bound in the bound complex can be measured using anodic stripping voltammetry and compared to the amount of metal-containing particles present in the removed electrolyte measured by anodic stripping voltammetry.
  • the determining step comprises voltammetry, including but not limited to, anodic stripping voltammetry, cathodic stripping voltammetry, adsorptive stripping voltammetry, square wave voltammetry, linear sweep voltammetry, staircase voltammetry, cyclic voltammetry, alternating current voltammetry, chronoamperometry, normal pulse voltammetry, differential -pulse voltammetry, or the like.
  • voltammetry including but not limited to, anodic stripping voltammetry, cathodic stripping voltammetry, adsorptive stripping voltammetry, square wave voltammetry, linear sweep voltammetry, staircase voltammetry, cyclic voltammetry, alternating current voltammetry, chronoamperometry, normal pulse voltammetry, differential -pulse voltammetry, or the like.
  • the methods described herein comprise exposing the analyte from the biological particle such that the analyte is available to form a bound complex.
  • the analyte-containing biological particle is lysed such that the analyte is released from the analyte-containing biological particle.
  • a lysing solution may be added to the analyte such that the analyte is released from the analyte-containing biological particle. Lysing solutions are described in more detail below.
  • the analyte-containing biological particle is lysed via mechanical agitation, heating, washing, and/or shearing of the analyte-containing biological particle (e.g., via ultrasonic agitation).
  • exposing analyte from the biological particle such that the analyte is available to form a bound complex comprises changing the pH, temperature, and/or ionic strength of the fluid comprising biological particle such that the analyte is available to form a bound complex.
  • Suitable capture substrates include woven pads, non-woven pads, non-magnetic resins, polymeric packing (e.g., polystyrene-divinylbenzene), and microfibers (e.g., electro spun microfibers).
  • the buffer solution comprises 50 mM sodium acetate, at least 1 mM and less than or equal to 5 mM zinc acetate, and at least 50 mM and less than or equal to 200 mM sodium chloride.
  • plasma separation of the sample is conducted prior to adding the sample to the capture substrates.
  • suitable methods such as centrifugation of filtration (e.g., a membrane based separator), for separating plasma from a sample (e.g., a whole blood sample).
  • the method may involve diagnosing the patient as not having hepatitis C, in embodiments in which the sample from the subject does not contain an HCV antigen and/or cAg.
  • a method involves identifying a patient, from two or more patients, as having or not having hepatitis C, by testing patient samples (e.g., whole blood samples) from the two or more patients according to one or more of the methods described herein. The method may involve determining the patient as having hepatitis C where the patient sample contains an HCV antigen and/or cAg, or determining that the patient does not have hepatitis C where the patient sample does not contain the HCV antigen and/or cAg.
  • the bound complex may be exposed to an electrolyte within a microfluidic device (e.g., within a channel of a microfluidic device).
  • an electric potential may be applied to at least a portion of a microfluidic device such that at least a portion of the metal from the metal-containing particles is oxidized, or such that at least a portion of the metal is deposited onto an electrode material within the microfluidic device (e.g., within a channel of a microfluidic device).
  • measuring current by changing a voltage to determine the amount of analyte present in the fluid occurs within the microfluidic device (e.g., within a channel of a microfluidic device).
  • the capture substrate may be present within, and/or a component of. the microfluidic device (e.g., a plurality of posts within a channel of the microfluidic device).
  • Virions attached to the capture substrates were lysed in a lysis solution containing detergents, and/or denaturants and/or reducing agents to release the core antigen, which was subsequently detected.
  • the lysis step opened up HCV particles to release the core antigen, monomerized the core antigen, inactivated the host-derived antibodies against the core antigen, and dissociated the core antigen from the interactions with blood components other than the antibody against the core antigen.
  • the wash buffer was composed of, for example, 0.1% Casein and 0.05% tween-20 in
  • Capture structures with attached virion particles were separated from blood solution by placing sample on magnet stage. All capture structures, including those with virions, are gathered near the magnet. Wash buffer was then added and the capture structures were resuspended by vortexing and pipetting up and down. The resuspended capture structures were then put back on the stage and the wash buffer was removed.
  • the wash buffer was composed of, for example, 50 mM sodium acetate, pH 5.6.
  • FIGs. 9A-9F show the effect of increasing the concentration of the electrolyte (NH 4 SCN), which resulted in changes in the peak area and shape.
  • 0.1M NH4SCN provided the largest peak area but distorted the shape of the peaks at high silver concentrations.
  • 0.5M NH4SCN, 1M NH4SCN, and 5 M NH4SCN were also tried and all of these decreased signal even farther.
  • the capture structures Prior to running the assay, can be, in some cases, coated with antibodies able to capture the analyte.
  • protocols for coating the particles with antibodies included EDC coupling to carboxylic acids present on the surface of the particles.
  • the size of the capture structure may be chosen as a balance between steric hindrance (e.g., smaller particles are generally more likely to successfully bind to an analyte on a large silver particle) and magnetic moment (e.g., larger particles generally have a higher magnetic moment and are more easily removed from solution with a magnet).
  • FIG. 2A An exemplary schematic of the above procedure is shown in FIG. 2A.

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  • Immunology (AREA)
  • Molecular Biology (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Engineering & Computer Science (AREA)
  • Hematology (AREA)
  • Electrochemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Biomedical Technology (AREA)
  • Urology & Nephrology (AREA)
  • Microbiology (AREA)
  • Cell Biology (AREA)
  • Food Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Biotechnology (AREA)
  • Spectroscopy & Molecular Physics (AREA)
  • Dispersion Chemistry (AREA)
  • Nanotechnology (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
  • Investigating Or Analyzing Materials By The Use Of Electric Means (AREA)

Abstract

L'invention concerne généralement des procédés de détermination et/ou de quantification d'un ou plusieurs analytes dans des fluides. Selon des modes de réalisation, un procédé consiste à introduire ou à exposer une pluralité de structures de capture conjuguées et une pluralité de particules conjuguées contenant du métal (par ex., de l'argent) à un fluide comprenant l'analyte de sorte que l'analyte se lie aussi bien à une structure de capture et à une particule contenant du métal (par ex., de l'argent) pour former un complexe lié. Le complexe lié peut alors être soumis à des conditions (par ex., des conditions électrochimiques) qui permettent la quantification de l'analyte sur la base de la quantité de particules contenant du métal présentes. Les procédés décrits ici peuvent être utiles à la détermination et à la quantification de concentrations relativement faibles d'analytes présents dans un échantillon d'un patient (par ex., une gouttelette de sang entier).
PCT/US2016/064539 2015-12-04 2016-12-02 Procédés de détermination d'analytes dans des fluides Ceased WO2017096114A1 (fr)

Applications Claiming Priority (2)

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US201562263399P 2015-12-04 2015-12-04
US62/263,399 2015-12-04

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WO2017096114A1 true WO2017096114A1 (fr) 2017-06-08

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2018119401A3 (fr) * 2016-12-22 2019-01-03 Daktari Diagnostics, Inc. Dispositifs et procédés pour déterminer un ou plusieurs analytes dans des fluides

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP4217741B1 (fr) * 2020-09-23 2025-04-16 Roche Diagnostics GmbH Procede de detection d'un analyte a l'aide de nanoparticules metalliques sur une electrode

Citations (5)

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Publication number Priority date Publication date Assignee Title
US20090107852A1 (en) * 2007-09-17 2009-04-30 Siometrix Corporation Self-actuating signal producing detection devices and methods
US20100075862A1 (en) * 2008-09-23 2010-03-25 Quanterix Corporation High sensitivity determination of the concentration of analyte molecules or particles in a fluid sample
US20110000785A1 (en) * 2007-10-31 2011-01-06 Raghbir Singh Bhullar Electrical patterns for biosensor and method of making
US20120279298A1 (en) * 2011-05-05 2012-11-08 Daktari Diagnostics, Inc. Conductive patterns and methods for making conductive patterns
US20130112572A1 (en) * 2011-11-04 2013-05-09 Ohmx Corporation Novel chemistry used in biosensors

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20090107852A1 (en) * 2007-09-17 2009-04-30 Siometrix Corporation Self-actuating signal producing detection devices and methods
US20110000785A1 (en) * 2007-10-31 2011-01-06 Raghbir Singh Bhullar Electrical patterns for biosensor and method of making
US20100075862A1 (en) * 2008-09-23 2010-03-25 Quanterix Corporation High sensitivity determination of the concentration of analyte molecules or particles in a fluid sample
US20120279298A1 (en) * 2011-05-05 2012-11-08 Daktari Diagnostics, Inc. Conductive patterns and methods for making conductive patterns
US20130112572A1 (en) * 2011-11-04 2013-05-09 Ohmx Corporation Novel chemistry used in biosensors

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2018119401A3 (fr) * 2016-12-22 2019-01-03 Daktari Diagnostics, Inc. Dispositifs et procédés pour déterminer un ou plusieurs analytes dans des fluides

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